Rat mouse gip total elisa

Manufactured by Merck Group

Sourced in United States, Japan

The Rat/Mouse GIP (total) ELISA is a laboratory equipment product designed for the quantitative determination of total GIP (Glucose-dependent Insulinotropic Polypeptide) in rat and mouse samples. This ELISA kit provides a reliable and accurate method for measuring GIP levels, which is a key hormone involved in the regulation of glucose homeostasis.

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7 protocols using rat mouse gip total elisa

Pancreatic Insulin and GIP Quantification

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Pancreatic tissue was homogenised in Krebs-Ringer buffer (pH 7.4) on ice. Tissue homogenate was extracted overnight in acid-ethanol (1.5% (vol./vol.) HCl in 75% (vol./vol.) EtOH). Tissue extracts were diluted 1:100 or 1:200 for insulin measurement. Diluted extracts were measured by HTRF Insulin Kit (Cisbio Bioassays, Codolet, France). Pancreatic insulin content was corrected by tissue weight for analysis. Tissue extracts were diluted 1:3 with Krebs-Ringer buffer for GIP measurement and protein content assay. Diluted extracts were measured by Rat/Mouse GIP (total) ELISA (Merck Millipore) and BCA protein assay (Sigma-Aldrich). Pancreatic GIP content was corrected by protein content for analysis.

Iida A., Seino Y., Fukami A., Maekawa R., Yabe D., Shimizu S., Kinoshita K., Takagi Y., Izumoto T., Ogata H., Ishikawa K., Ozaki N., Tsunekawa S., Hamada Y., Oiso Y., Arima H, & Hayashi Y. (2016). Endogenous GIP ameliorates impairment of insulin secretion in proglucagon-deficient mice under moderate beta cell damage induced by streptozotocin. Diabetologia, 59, 1533-1541.

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Measuring Glucose, GIP, and GLP-1 in Mice

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Blood glucose levels were measured using an Antsense Blood Glucose Meter (Horiba, Kyoto, Japan). Plasma GIP and insulin were measured using Rat/Mouse GIP (total) ELISA (Merck Millipore, Darmstadt, Germany) and Mouse Insulin ELISA Kit (Morinaga Institute of Biological Science, Kanagawa, Japan) as previously reported [21 (link)]. Blood samples for measurements of biologically intact GLP-1 and GIP were collected using BD P800 tubes (Becton Dickinson, Franklin Lakes, NJ, USA). Plasma levels of biologically intact GLP-1 and GIP were evaluated using the following immunoassays according to manufacturers’ instructions: GLP-1, Active GLP-1 (ver. 2) Kit (Meso Scale Discovery, Rockville, MD, USA) and GIP, Mouse GIP, Active form Assay Kit (Immuno-Biological Laboratories, Gunma, Japan).

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Metabolic Biomarker Profiling in Animal Model

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Blood samples collected after a 6-hour fasting were used for analysis. Hemoglobin A1c (HbA1c) levels were measured using an A1C Now-Plus (Bayer, Frankfurt, Germany) before sacrificing the animals. Plasma levels of glucose, total cholesterol, high-density lipoprotein (HDL) cholesterol, and triglyceride were measured by enzymatic methods (WAKO, Osaka, Japan). Plasma levels of insulin, active glucagon-like peptide-1 (GLP-1), and total glucose-dependent insulinotropic polypeptide (GIP) were determined by enzyme-linked immunosorbent assay (ELISA) (ultrasensitive mouse insulin ELISA kit from MORINAGA, Kanagawa, Japan; GLP-1 (active) ELISA and Rat/Mouse GIP (total) ELISA from EMD Millipore (MA, USA). Blood pressure and pulse were measured on the day of sacrifice in the fasting state using the tail-cuff method (Model MK-2000ST, Muromachi Kikai Co., Ltd., Tokyo, Japan) [5 (link)–7 (link), 9 (link), 10 (link)].

Terasaki M., Hiromura M., Mori Y., Kohashi K., Kushima H., Ohara M., Watanabe T., Andersson O, & Hirano T. (2017). Combination Therapy with a Sodium-Glucose Cotransporter 2 Inhibitor and a Dipeptidyl Peptidase-4 Inhibitor Additively Suppresses Macrophage Foam Cell Formation and Atherosclerosis in Diabetic Mice. International Journal of Endocrinology, 2017, 1365209.

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Incretin Levels and Atherosclerosis in Mice

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Glycated hemoglobin (HbA1c) levels were measured through a cut in the tail vein using the quick test A1C Now Plus (Bayer, Frankfurt, Germany) before sacrificing the animals. The blood samples were collected after a 6-h fast. The plasma levels of glucose, total cholesterol, high-density lipoprotein (HDL) cholesterol, and triglyceride were measured by enzymatic methods (WAKO, Osaka, Japan). We had previously reported that dipeptidyl peptidase-4 inhibitor elevated active glucagon-like peptide-1 (GLP-1) and total glucose-dependent insulinotropic polypeptide (GIP), that prevented the acceleration of atherosclerosis by suppressing foam cell formation in Apoe^−/− mice [22 (link), 23 (link)]. Therefore, we measured the endogenous incretin levels in this study. The plasma levels of insulin, active GLP-1, and total GIP were determined by enzyme-linked immunosorbent assay (ELISA) (Ultra-sensitive mouse insulin ELISA kit from Morinaga, Kanagawa, Japan; GLP-1 (active) ELISA and Rat/Mouse GIP (total) ELISA from EMD Millipore (Billerica, MA, USA).

Terasaki M., Hiromura M., Mori Y., Kohashi K., Nagashima M., Kushima H., Watanabe T, & Hirano T. (2015). Amelioration of Hyperglycemia with a Sodium-Glucose Cotransporter 2 Inhibitor Prevents Macrophage-Driven Atherosclerosis through Macrophage Foam Cell Formation Suppression in Type 1 and Type 2 Diabetic Mice. PLoS ONE, 10(11), e0143396.

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Serum Biomarker Analysis in Animal Model

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A total of one day after the last behavioral tests, animals were killed by cervical decapitation previous anesthesia. Blood was extracted by cardiac puncture and serum was obtained and stored a −70 °C until analysis. Serum parameters including glucose, triglyceride (TG), and total cholesterol were measured using an enzymatic-photometric assay with a Cobas c111 analyzer (Roche Applied Science). Insulin (ELISA kit Alpco Diagnostics, Salem, MA, USA), LPS (Cloud-Clone Corp, Houston, TX, USA), leptin (rat leptin ELISA kit, St Charles, MO, USA) and GIP (Merck-Millipore rat/mouse GIP (total) ELISA, Bellerice, MA, USA) were measured by ELISA.

Syeda T., Sánchez-Tapia M., Orta I., Granados-Portillo O., Pérez-Jimenez L., Rodríguez-Callejas J.D., Toribio S., Silva-Lucero M.D., Rivera A.L., Tovar A.R., Torres N, & Perez-Cruz C. (2021). Bioactive Foods Decrease Liver and Brain Alterations Induced by a High-Fat-Sucrose Diet through Restoration of Gut Microbiota and Antioxidant Enzymes. Nutrients, 14(1), 22.

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Metabolic Biomarkers in Animal Model

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Fasting blood glucose and hemoglobin A1c (HbA1c) levels were measured through a cut in the tail vein using a dextrometer (Glutest Sensor; Sanwa Kagaku, Aichi, Japan) and A1CNow (Bayer Inc, Leverkusen, Germany) before sacrificing the animals. Plasma levels of total cholesterol and triglycerides were measured by using the enzymatic colorimetric method (Wako). Plasma levels of insulin and total GIP were determined using enzyme-linked immunosorbent assays (ELISAs) (Ultra-sensitive mouse insulin ELISA kit; Morinaga, Kanagawa, Japan; Rat/Mouse GIP (total) ELISA, EMD Millipore).

Hiromura M., Mori Y., Kohashi K., Terasaki M., Shinmura K., Negoro T., Kawashima H., Kogure M., Wachi T., Watanabe R., Sato K., Kushima H., Tomoyasu M., Nakano Y., Yamada Y., Watanabe T, & Hirano T. (2016). Suppressive Effects of Glucose-Dependent Insulinotropic Polypeptide on Cardiac Hypertrophy and Fibrosis in Angiotensin II-Infused Mouse Models. Circulation journal : official journal of the Japanese Circulation Society, 80(9).

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Measurement of Metabolic Biomarkers in Rats

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The levels of blood HbA1c were measured using a commercial kit based on the latex-agglutination assay (RAPIDIA ® Auto HbA1c-L; Fujirebio Inc.). Plasma TAG, cholesterol, free fatty acids and glucose levels were measured using enzyme assay kits (Triglyceride E-Test, Cholesterol E-Test, NEFA C-Test and Glucose CII-Test, respectively; Wako Pure Chemicals). Plasma levels of insulin, glucagon, glucose-dependent insulinotropic polypeptide (GIP) and GLP-1 and serum levels of fatty acidbinding protein 4 (FABP4) and SAP were measured using commercial ELISA kits (rat insulin ELISA; Shibayagi Co. Ltd, rat/mouse GIP total ELISA; EMD Millipore Co., rat glucagon ELISA kit and rat GLP-1 ELISA kit; Wako Pure Chemicals, CircuLex Rat FABP4/A-FABP ELISA kit; CycLex Co. Ltd, Serum Amyloid P Rat ELISA kit; Abcam). Glycogen levels in the liver and muscle were determined according to the method of Lo et al. (37) .

Shirouchi B., Nagao K., Umegatani M., Shiraishi A., Morita Y., Kai S., Yanagita T., Ogawa A., Kadooka Y, & Sato M. (2016). Probiotic Lactobacillus gasseri SBT2055 improves glucose tolerance and reduces body weight gain in rats by stimulating energy expenditure. The British journal of nutrition, 116(3).

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